全反式维甲酸对环孢素诱导的肾小球系膜细胞增殖与凋亡的影响及其机制

陈旭; 吴建华; 郭乃凤; 范亚平; 陈晓岚

doi:10.3969/j.issn.1674-7445.2017.06.011

全反式维甲酸对环孢素诱导的肾小球系膜细胞增殖与凋亡的影响及其机制

doi: 10.3969/j.issn.1674-7445.2017.06.011

226001 江苏南通，南通大学附属医院肾内科

基金项目:

江苏省自然科学基金 BK2008185

南通市科技项目 MS32015018

详细信息

通讯作者:
陈晓岚, Email: chenxl8448@sina.com

中图分类号: R617, R392.4, R692
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- 文章访问数: 112
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- PDF下载量: 5
- 被引次数: 0
出版历程
- 收稿日期: 2017-08-30
- 网络出版日期: 2021-01-19
- 刊出日期: 2017-11-15

Effect and mechanism of all-trans retinoic acid on cyclosporin-induced proliferation and apoptosis of glomerular mesangial cells

Department of Nephrology, the Affiliated Hospital of Nantong University, Nantong 226001, China

More Information

Corresponding author: Chen Xiaolan, Email: chenxl8448@sina.com

摘要

摘要: 目的探讨全反式维甲酸（ATRA）对环孢素（CsA）诱导的大鼠肾小球系膜细胞增殖及凋亡的影响及机制。方法采用不同剂量ATRA作用于不同剂量CsA诱导下的肾小球系膜细胞。采用MTT比色法检测细胞增殖，Hoechst 33258荧光染色观察细胞凋亡形态，后用流式细胞仪检测细胞凋亡率，免疫荧光法检测线粒体促凋亡的Smac蛋白的表达。结果与对照组比较，CsA剂量为0.5 μg/mL及以上可抑制细胞増殖，CsA剂量为1.0 μg/mL及以上可诱导细胞凋亡，且Smac蛋白的表达随CsA剂量的增加而增加，具有剂量和时间依赖的特征（均为P＜0.05）。而与CsA组比较，CsA+ATRA处理组对细胞增殖的抑制作用更为明显，而加入ATRA可呈剂量依赖性抑制CsA诱导的肾小球系膜细胞凋亡及Smac蛋白的表达（均为P＜0.05）。结论 CsA可抑制肾小球系膜细胞增殖，诱导其凋亡并刺激Smac蛋白的表达。ATRA可以抑制CsA诱导的肾小球系膜细胞凋亡，且可能是通过Smac信号通路介导的。
- 全反式维甲酸（ATRA） /
- 环孢素（CsA） /
- 系膜细胞，肾小球 /
- Smac蛋白 /
- 排斥反应 /
- 细胞增殖 /
- 凋亡 /
- 肾毒性
Abstract: Objective To investigate the effect and mechanism of all-trans retinoic acid (ATRA) on the cyclosporin (CsA)-induced proliferation and apoptosis of glomerular mesangial cells in rat models. Methods The glomerular mesangial cells induced by different doses of CsA were treated with different doses of ATRA. MTT assay was carried out to detect cell proliferation. Hoechst 33258 fluorescent staining was adopted to observe the morphology of the apoptotic cells. Flow cytometry was conducted to detect the cellular apoptosis rate. Immunofluorescent staining was employed to quantitatively measure the expression level of mitochondria-derived pro-apoptotic Smac protein. Results Compared with the control group, administration of CsA at a dose of 0.5 μg/mL and above could suppress cellular proliferation, and use of CsA at a dose of 1.0 μg/mL and above could induce cellular apoptosis. The expression level of Smac protein was significantly up-regulated by CsA administration with a dose and time dependence (all P < 0.05).Compared with the CsA group, combined administration of CsA and ATRA exerted a more significant inhibitory effect on cellular proliferation. Supplement of ATRA could significantly inhibit glomerular mesangial cellular apoptosis induced by CsA and down-regulate the expression of Smac protein with a dose dependence (both P < 0.05). Conclusions CsA can inhibit cellular proliferation, induce cellular apoptosis and up-regulate the expression of Smac protein of glomerular mesangial cells. ATRA is capable of suppressing glomerular mesangial cellular apoptosis induced by CsA, which is probably mediated by the Smac signaling pathway.
- All-trans retinoic acid (ATRA) /
- Cyclosporin (CsA) /
- Mesangial cell, glomerulus /
- Smac protein /
- Rejection reaction /
- Cell proliferation /
- Apoptosis /
- Renal toxicity

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图 1 ATRA对CsA所致各组肾小球系膜细胞凋亡形态学及荧光强度的影响（免疫荧光，×400）

A ~ I图分别为A ~ I组的肾小球系膜细胞凋亡的荧光图。J图中，与A组比较，^aP＜0.05；与E组比较，^bP＜0.05；与F组比较，^cP＜0.05

Figure 1. Effect of ATRA on apoptotic morphological characteristics and fluorescence intensity of glomerular mesangial cell induced by CsA

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图 2 ATRA及CsA对肾小球系膜细胞凋亡率的影响

A ~ I图分别为A ~ I组肾小球系膜细胞凋亡的流式细胞图。J图中，与A组比较，^aP＜0.05；与E组比较，^bP＜0.05；与F组比较，^cP＜0.05

Figure 2. The effect of ATRA and CsA on apoptosis rate of glomerular mesangial cell

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图 3 ATRA及CsA对肾小球系膜细胞Smac蛋白表达的影响（免疫荧光，×400）

A ~ I图分别为A ~ I组肾小球系膜细胞Smac蛋白的免疫荧光图片。J图中，与A组比较，^aP＜0.05；与E组比较，^bP＜0.05；与F组比较，^cP＜0.05

Figure 3. Effect of ATRA and CsA on expression of Smac protein of glomerular mesangial cell

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表 1 ATRA和CsA对肾小球系膜细胞增殖的影响

Table 1. Effect of ATRA and CsA on proliferation of glomerular mesangial cell

(x±s)
组别	n	A₅₇₀值
组别	n	24 h	48 h	72 h
A组	3	0.672±0.028	0.708±0.016	0.723±0.035
B组	3	0.635±0.020^a	0.592±0.029^a,d	0.549±0.018^a,d
C组	3	0.587±0.031^a	0.523±0.024^a,d	0.459±0.033^a,d
D组	3	0.471±0.025^a	0.416±0.019^a,d	0.362±0.023^a,d
E组	3	0.418±0.026^a	0.365±0.022^a,d	0.314±0.030^a,d
F组	3	0.668±0.024	0.702±0.018	0.718±0.030
G组	3	0.384±0.019^a,b,c	0.316±0.028^a,b,c,d	0.253±0.025^a,b,c,d
H组	3	0.313±0.036^a,b,c	0.247±0.016^a,b,c,d	0.182±0.014^a,b,c,d
I组	3	0.249±0.027^a,b,c	0.175±0.030^a,b,c,d	0.116±0.023^a,b,c,d
与A组比较，^aP＜0.01；与E组比较，^bP＜0.05；与F组比较，^cP＜0.05；与同组24 h比较，^dP＜0.05

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